Neem Formulations Research Articles

Comparative Toxicity of Neem and Peppermint Oils Nano Formulations against Agrotis ipsilon (Hufn.) Larvae (Lepidoptera: Noctuidae)

Applications of nanotechnology in agriculture will result in the development of efficient and potential approaches towards the management of insect pests. The toxicity effects of four essential oils peppermint, thyme, camphor and sage oils were tested against the fourth instar larvae of Agrotis ipsilon to select the most effective essential oil to be converted to the nano form. According to the results obtained, peppermint oil was the most toxic compound, which has been used in the present investigation compared with neem oil. The toxicity of bulk and nano- formulations of neem and pepper mint oils were tested against 2nd and 4th instar larvae of A. ipsilon under laboratory conditions of 25±2 °C& 65 -70 % R.H.relative humidity The results show that the LC50 value (the concentration used which kill 50% of the tested individuals)of loaded neem or pepper mint were lower (0.62 and 36.47 ppm) compared with neem or pepper mint oil nano-emulsion and bulk neem for the second larval instar. The different formulations of neem are more potent than in case of peppermint oil, as LC50 and LC90 values were significantly lower.The same trend was found concerning the 4th larval instar. Age of treated larvae had a detrimental effect on the response to the compounds tested. It was noticed that the younger larvae were much more sensitive to the prepared compounds compared to the older ones. The least LC50 value for loaded neem nano-emulsion was 6.68 ppm compared with the highest value for bulk neem oil (16.68 ppm ). Also, LC90 values followed the same trend as in case ofLC50. Again, the toxicity of loaded peppermint oil had the most insecticidal activity as expressed by the lowest LC50 value (51.9 ppm) with more insecticidal effect than the bulk(125.43 ppm) or nano-emulsion (85.43 ppm). The present results indicated that these novel systems could be used in integrated pest management program for A. ipsilon control.

Laboratory Evaluation of Neem formulation bioactivity against Crocidolomia pavonana F. larvae

This study aims to evaluate the bioactivity of the formulation of Neem 50 EC against Crocidolomia pavonana larvae in the laboratory. The study using six treatments and three replications. The treatment consisted of the formulation of neem 50 EC at concentrations of 0.3%, 0.5%, 0.8%, 1.3%, 2.3%, and control. The treatment was done by leaf feeding method and tested on C. pavonana instar II. The results of the research showed that the formulation of neem 50 EC at concentration of 2.3% caused mortality of C. pavonana larvae with mortality rate of 95% and has LC50 value aqual to 0.83%. Neem formulation can inhibit the development of C. pavonana larvae from instar I to instar IV, decrease food consumption, and decrease the dry weight of C. pavonana larvae instar IV.Keywords: Azadirachta indica, Bioactivity, Crocidolomia pavonana, neem formulation

Testing The Effect of Temperature, Storage Time, The Residual Test of Neem Oil Formulation (Azadirachta indica A. JUSS) and Bitung Formulation (Baringtonia asiatica) to Its Toxicity Against Large Cabbage Heart Caterpillar (Crocidolomia pavonana F)

This study aims to determine the effect of temperature and storage time of neem formulation and residual test of neem formulation and bitung formulation to its toxicity on C. pavonana larvae. The formulations tested on the storability and temperature test using the newly prepared, neem formula has been stored for 19 months at room temperature 24.6-31,7 C, and stored in low temperature (4 degree Celsius) as well as controls. The toxicity of the neem oil formulation was tested at concentrations of 1%, 2%, and 3%. Residual testing was performed by spraying and the residual test samples were taken on days 0,1,2,3,5,7,9,11, and 13 days after application of insecticide spraying. The results showed that storage at low temperatures (temperature 4 degree Celsius) for 19 months can maintain the toxicity of the neem formulation to C. pavonana larvae. while storage at room temperature for 19 months resulted in a decreased toxicity. The residual activity of the neem formulations of concentration 0.8% and 2.4% ceased to cause the death C. pavonana larvae at the residual age of 13.34 days and 14.66 days. The activity of Bitung formulation concentration of 1.1% and 2.8% ceased to causing death C. pavonana larvae at the residual age of 11.33 days and 16.26 days.Keyword: Azadirachta indica, Baringtonia asiatica, Crocidolomia pavonana, Residue test, Storability test, Temperatures, Toxicity.

Efficiency of Nano-formulations of Neem and Peppermint Oils on the Bionomics and Enzymatic Activities of <i>Agrotis ipsilon</i> Larvae (Lepidoptera: Noctuidae)

Nanotechnology is considered to be a development way to gain efficient and potential approaches towards pest control. The effect of both neem and peppermint oils as bulk, nano and loaded nano-emulsions have been studied against the different biological aspects of the 2nd larval instar of Agrotis ipsilon (Hufn.) under laboratory conditions. Their effects on enzymatic activities were investigated for the 2nd and 4th larval instars. The effects of adding the different formulations of either neem or peppermint oils to artificial diet of 2nd instar larvae show significant elongation of the larval duration; percentage mortalities were increased as well as larval malformations. It was recorded that nano-formulations (emulsion or loaded) significantly decreased the pupal weight and significantly increased the pupal duration, percentage pupal mortalities and pupal malformation. Adult longevity show insignificant effect, while female fecundity and percentage egg fertility show significant decrease in comparison with the control. Results of enzymatic activities show marked effects of the three formulations of either neem or peppermint oil. Significant inhibitions were recorded for amylase, invertase, trehalase, protease and alkaline phosphatase. However, significant increase in the activities of cuticle phenoloxidase and chitinase were recorded. Marked variations were recorded between nano-formulations and the bulk form.

Formulation and evaluation of Neem (Azadirachta indica A. Juss) seed oil suppositories

Background: Neem seed is very vital because of its rich lipid content and bitter constituents. Aims: This study was designed to provide a scientific rationale for the preparation and use of Neem seed oil as suppositories using dika fat (DF), and macrogol (MG), as bases. Materials and Methods: The suppositories which were prepared by fusion method using a pre-calibrated mould, were characterized using parameters like appearance, crushing strength, weight variation, melting point, pH, liquefaction time and in-vitro release according to standard procedures. Results: Results show that, the suppository strengths were in the order; bland, MG (25.00 ± 1.50N) > DF (12.90 ± 0.72 N), while those containing medicaments were NSM1 (20.00 ± 1.92) > NSD1 (12.90 ± 0.94) > NSM2 (12.70 ± 1.24 N) > NSD2 (10.00 ± 1.35 N). The pH of medicated formulations were NSM1 (6.50 ± 0.01), NSM2 (6.54 ± 0.03) > NSD1 (5.73 ± 0.04) and NSD2 (5.07 ± 0.03). Melting point values show that, macrogol base had mean values of 36.80 °C ± 0.62 and 36.40 °C ± 0.46 for NSM1 and NSM2 respectively, whereas, those with DF gave an average melting point values of 32.10 °C ± 0.87 and 30.90 °C ± 0.79 for NSD1 and NSD2 respectively. Conclusion: Results obtained showed that suppositories prepared with Macrogol (MG) base exhibited better physicochemical properties than Dika fat (DF) base suppositories, therefore water soluble bases may be bases of choice in the delivery of neem seed oil.

Simultaneous determination of five azadirachtins in the seed and leaf extracts of Azadirachta indica by automated online solid-phase extraction coupled with LC\u2013Q-TOF\u2013MS

Neem (Azadirachta indica) extract is well-known as a natural pesticide for the control of agricultural pests. Azadirachtin A and its structural analogues are considered as active compounds. However, the amounts of azadirachtins varies in neem extracts, providing a variety of insecticidal activities. In this study, a novel method of automated online solid-phase extraction coupled with liquid chromatography/quadrupole-time-of-flight mass spectrometry (SPE-LC–Q-TOF–MS) was developed and validated for simultaneous quantification of five azadirachtins (azadirachtins A, B, D, H and I) in seed and leaf extracts of A. indica. Different experimental parameters (such as SPE cartridge, injection volume and washing step) were optimized. The optimized SPE-LC–Q-TOF–MS method showed good recovery (82.0–102.8%), linearity (r2 ≥ 0.9991) and precision (0.83–4.83%). The limit of detections (LODs) for the five analytes ranged from 0.34 to 0.76 ng mL−1. The validated method was successfully applied for determination of the analytes in the neem leaves and seeds from different locations and a neem formulation. The online SPE-LC–Q-TOF–MS method was found to be a simple, precise and accurate and can be used as a powerful tool for quality control of neem extracts or its formulations.

Effect of oil cakes and garlic aqueousbased formulations of trichoderma viride on management of meloidogyne incognita in chilli

Abstract Root-knot nematode, Meloidogyne incognita (Kofoid and White) is a major threat to chilli (Capsicum annum) cultivation, by forming root galls and subsequent wilting. These nematodes live in soil, roots debris and reported surviving in other Solanaceae crops. Chemical application of nematicides cause impact on environment, therefore biocontrol using antagonistic fungi is desired to tackle this problem. This research is aimed to evaluate the effect of Trichoderma viride and its formulations on management of M. incognita. Oil cakes such as neem (Azadiracta indica), gingelly (Sesamum indicum), Mahua (Madhuca longifolia) and garlic (Allium sativum) extracts were used to evaluate the growth and sporulation of T. viride. Chilli variety MI-2 was used. Among the four preparations, neem oil cake recorded high spore yield of 1.75 ×107 spores/ml. Similarly gingelly oil cake and garlic produced the spore yields of 1.57 ×107 spores/ml and 1.368 ×107 spores/ml, respectively. Comparatively Mahua oil cake obtained low spore yield of 7.87×106 spores/ml. Plant growth was significant (P<0.05) in the application of neem oil cake formulation (30.42 cm). Extent of galling was significantly low with garlic (9.00) and neem (9.25). These results confirmed that the formulations of neem, gingelly oil cake extracts and garlic aqueous extract stimulated the chilli plant growth, productivity and reduced the nematode infestation. Above all, neem oil cake and garlic are the best formulations that can be used to manage M. incognita

EVALUATION OF BT LIQUID FORMULATIONS AGAINST GRAM POD BORER, HELICOVERPA ARMIGERA (HUBNER) AND SPOTTED POD BORER, MARUCA VITRATA (GEYER) IN PIGEONPEA

A field experiment was conducted during 2012-15 at Regional Agricultural Research Station, Lam, Guntur district, Andhra Pradesh to evaluate the efficacy of Bt liquid formulations and other biopesticides against gram pod borer, Helicoverpa armigera and spotted pod borer, Maruca vitrata in pigeonpea. There were nine treatments [2 strains of Bt liquid formulations each at two doses, two doses of Beauveria bassiana, neem formulation (azadirachtin 1500 ppm), and chemical check] including untreated control. Pooled analysis of three years data revealed that two sprays of NBAII BtG4 @ 2% at fortnight interval significantly superior over other treatments in suppressing the larval population of H. armigera (3.2 larvae / plant) and M. vitrata (5.9 larvae / plant) on pigeonpea and recorded minimum pod damage (3.7 and 11.4%, respectively) with maximum yield (1565 kg/ha). Further, it was also revealed that all Bt liquid formulations and B. bassiana were safe to natural enemy population viz., spiders and coccinellids existing in pigeonpea ecosystem. It is suggested to utilize biopesticides as ecofriendly insecticides for pigeonpea production.

Laboratory Evaluation of Neem formulation bioactivity against Crocidolomia pavonana F. larvae

Laboratory Evaluation of Neem formulation bioactivity against Crocidolomia pavonana F. larvae

Colour and spreadability of Neem (Azadirachta Indica A. juss) ointment and cream formulations

Herbal plants are a major source of raw material for traditional medicines. Recently there has been an increase of interest to study the therapeutic potential of herbal plants as herbal care products. In this study, a preliminary study on the formulation of neem (Azadirachta Indica) ointment and cream have been conducted. The neem leaves were extracted and formulated into ointment and cream. The raw neem extract is added into the ointment and cream bases at four different concentrations (0% w/w, 0.5% w/w, 1% w/w and 2% w/w) and stored at three different storage temperatures (4°C, 25°C and 45°C). The semambu ointment and cream formulated were evaluated in terms of their colour and spreadability. It has been found that the extract content and storage temperature influence the colour and spreadability of the formulated neem ointment and cream.

Field trials of formulated neem seed oil at different rates and frequencies on pigeonpea flower blister beetle, Mylabris pustulata Thunberg in Owerri Rainforest Zone, Nigeria

In this study, the efficacy of Formulated Neem Seed Oil (FNSO) on the population of blister beetle, Mylabris pustulata Thunberg (Coleoptera: Meloidae) was evaluated. The data were recorded from beginning of May 2009 and 2010 planting seasons. The experiment was laid out in 3×5 factorial comprising three rates of neem seed oil, 4.2 L ha–1, 8.3 L ha–1,12.5 L ha–1 with Control (0 L ha–1), and synthetic pyrethroid, cypermethrin (1.5L ha–1) as checks plots. There were also three intervals of application: once a week, once in two weeks, and once in three weeks. Results showed that application of F-NSO at higher dosage of 12.5L ha–1 and at four regime spraying intervals of once a week significantly (P<0.05) reduced the population of blister beetles on flowers of short duration pigeonpea cultivar (ICPL 84023). This effective application dose of 12.5L ha–1 at four regime spraying intervals of once a week can be an ideal Integrated Pest Management tool for the management of M. pustulata in Rainforest Zone of Southeastern, Nigeria.

Neem extract formulations to control Diabrotica speciosa (Coleoptera: Chrysomelidae) larvae in maize

Diabrotica speciosa larvae have been considered to be one of the major subterranean pests of several crops, and its most susceptible host is maize (Zea mays). This study aimed to evaluate the effects of oil-formulated neem extract and lignin-microencapsulated neem extract for D. speciosa larvae control. Effects of applications of neem formulations on the growth of maize plants were also assessed. In the first assay, 0.25, 0.5, 1.0, 2.0, and 4.0 mL of oil-formulated neem extract were applied per 25 g soil and were compared with the insecticide fipronil WG and deionized water. In the second assay, lignin-microencapsulated neem extract was applied at 160, 80, 40, 20 and 10 mg per 25 g soil, and compared with the same controls. Both neem formulations were as efficient as the insecticide fipronil WG in the control of D. speciosa larvae. The evaluated doses of neem formulations did not interfere on maize plants growth. Results of this study permit concluded that, under laboratory conditions, oil-formulated neem extract and lignin-microencapsulated neem extract formulations are promising for controlling D. speciosa larvae in maize, and might benefit sustainable pest management.

Resistance of important bean genotypes to the Mexican bean beetle [Zabrotes subfasciatus (Bohemann)] during storage and its control with chemical synthetic and botanical insecticides

During storage, beans can be infested with many insect-pests including Zabrotes subfasciatus, a key pest of these crops. This study aimed to identify bean genotypes demonstrating antixenosis and/or antibiosis to Z. subfasciatus and to test their integration with chemical control. The ultimate goal of assessment was to distinguish whether genotypic and insecticidal factors can provide effective beetle control. The tested genotypes were (a) Phaseolus vulgaris group: CCB, BSCB, BCB and PCB; (b) Vicia faba group: YBB, WBB and SBB; and (c) Vigna unguiculata group: C and GC. Initial assays were run to select genotypes (without insecticide treatment) that would be further tested with insecticides. Final assays included genotypes with varying degree of antibiosis and antixenosis treated with a neem formulation (Natuneem) and distilled water (control) plus deltamethrin (Decis) which latter was used only in the final antibiosis assay. The insecticides were used at the rates of 3 and 0.1 mL of Natuneem and Decis, respectively, per 30 mL of distilled water. There were no differences in preference of Z. subfasciatus adults among non-treated genotypes (initial assays), although neem-treated genotypes altered the preference and reduced infestation from 40.54-100% (final assays). In antibiosis tests, oviposition and density of emerged adults were reduced among C and SBB, and SBB also reduced the weight of emerged adults. Insecticides reduced oviposition in 53-100% and yielded half to five-fold fewer emerging insects weighting 35%-40% less in antibiotic genotypes. SBB was the most antibiotic genotype and this and other genotypes possessing antibiosis had a synergistic effect with neem or deltamethrin.

Novel and environmental friendly approach; Impact of Neem (Azadirachta indica) gum nano formulation (NGNF) on Helicoverpa armigera (Hub.) and Spodoptera litura (Fab.)

The future of this study was to prepare a natural pesticide which will not harm the environment and yet control pests. Neem gum nano formulation (NGNF), a novel biopesticide prepared from the Neem gum extract (Azadirachta indica) (NGE) was evaluated for its antifeedant, larvicidal and pupicidal activities against Helicoverpa armigera (Hub.) and Spodoptera litura (Fab.) at 100ppm. The NGNF showed significant (100%) antifeedant, larvicidal and pupicidal activities against H. armigera and S. litura. The LC50 values of 10.20, 12.49 and LC90 values of 32.68, 36.68ppm on H. armigera and S. litura, respectively at 100ppm. The NGNF treatments showed differences in the activities of detoxifying enzymes, carboxylesterases, glucosidases and glutathione S-transferases in the larval gut. Earthworm toxicity illustrated that 6.25ppm of chemical insecticides (cypermethrin) varied widely in their contact toxicities compared to 100ppm of NGNF and control in both contact filter paper and artificial soil test. The NGNF were characterized and confirmed by FTIR, XRD, SEM and EDX analysis. Ten compounds were identified from the Neem gum extract (NGE) by Gas Chromatography-Mass Spectrometry (GC–MS) analysis. The major compounds were fatty acids like Hexadecanoic acid, oleic acid, and ricinoleic acid. NGNF could be used as an agent to prepare novel bio-pesticides formulations.

Nanocapsules Containing Neem (Azadirachta Indica) Oil: Development, Characterization, And Toxicity Evaluation

In this study, we prepared, characterized, and performed toxicity analyses of poly(ε-caprolactone) nanocapsules loaded with neem oil. Three formulations were prepared by the emulsion/solvent evaporation method. The nanocapsules showed a mean size distribution around 400 nm, with polydispersity below 0.2 and were stable for 120 days. Cytotoxicity and genotoxicity results showed an increase in toxicity of the oleic acid + neem formulations according to the amount of oleic acid used. The minimum inhibitory concentrations demonstrated that all the formulations containing neem oil were active. The nanocapsules containing neem oil did not affect the soil microbiota during 300 days of exposure compared to the control. Phytotoxicity studies indicated that NC_20 (200 mg of neem oil) did not affect the net photosynthesis and stomatal conductance of maize plants, whereas use of NC_10 (100:100 of neem:oleic acid) and NC_15 (150:50 of neem:oleic acid) led to negative effects on these physiological parameters. Hence, the use of oleic acid as a complement in the nanocapsules was not a good strategy, since the nanocapsules that only contained neem oil showed lower toxicity. These results demonstrate that evaluation of the toxicity of nanopesticides is essential for the development of environmentally friendly formulations intended for applications in agriculture.

Characterization and Testing of Fine Powder Formulation of Whole Neem Fruits

Azadirachtin (Aza) is a key ingredient of neem-based pesticides. However, use of neem pesticides is limited due to storage instability of Aza. In this work, freeflowing fine powder of whole dry neem fruits (powder neem formulation, PNF) is developed without separately extracting Aza. The optimal particle size was found to be −44 + 60 mesh. PNF is characterized by Fourier transform infrared, X-ray diffraction, Brunauer- Emmett-Teller surface area, particle-size distribution and scanning electron microscope. Stability of Aza was found to be improved and it was assessed by studying the effect of particle size, temperature, UV light exposure and release study in buffered and natural water samples.

Comparative efficacy and phytotoxicity evaluation of biopesticides, insecticides and Neem formulation against leaf folder (Cnaphlocrocis medinalis guenee) on paddy

Effort were taken to compare the efficacy and phytotoxicity of bioagent B.bassiana Neem formulation and insecticide. Out of six treatment Monocrotophos 36 per cent SL @ 625 ml/ha and Beauveria bassiana 1.15 per cent WP (1x10 8 cfu/g min.) treatments @ 3000 and 2500 g/ha were effective to reducing leaf folder larval population on paddy crop and to increase the grain yield. All the treatments were non-phytotoxic to paddy crop and non-toxic to natural enemies in both the year. Beauveria bassiana 1.15 per cent WP applied @ 2500 g/ha dose was optimum to control leaf folder and to increase the yield. Based on the results of bioefficacy and grain yield, use of Beauveria bassiana 1.15 per cent WP @ 2500 g/ha is suggested for the effective management of leaf folder larvae on paddy crop.

Integrated approach for the management of white grub and collar rot in groundnut (Arachishypogaea L.) in South West Haryana

Field experiments were conducted at Chaudhary Charan Singh Haryana Agricultural University, Regional Research Station, Bawal (Rewari), Haryana during Kharif seasons of 2011 and 2012. Fifteen treatments including three insecticides, one fungicide, three antagonists, one amendment and one neem formulation were evaluated in sole and combination treatments as seed dresser whereas chlorpyriphos 20 EC and poultry manure were incorporated in soil to find out effective combination for control of collar rot and white grub of groundnut. Minimum disease incidence 0.3 and 0.7 per cent and white grub infestation 1.2 and 1.5 per cent was recorded during 2011 and 2012 with seed treatment of chlorpyriphos 20 EC @ 25 ml/kg seed + PM 4 @ 10 ml/kg seed + captan 3g/kg seed followed by imidacloprid 17.8 SL @ 7 ml/kg seed and chlorpyriphos 20 EC @ 25 ml /kg seed + Bacillus thuriengensis@ 10 ml/ kg seed + captan 3 g/kg seed. The yield was significantly (at 5% level) low (7.10 q/ha during 2011 and 6.8 q/ha during 2012) in control as compared to chlorpyriphos 20 EC + PM-4 + captan treatment (20.20 q/ha during 2011 and 19.80 q/ha during 2012) which ranked at first place. The seed treated with imidacloprid 17.8 SL 7 ml /kg seed also produced good yield in both the years and was rated as second best treatment followed by chlorpyriphos + Bacillus thurengiensis+ captan. The results obtained herein provide the foundation for an effective and sustainable programme of management for groundnut white grub and collar rot. Integrated approach having chlorpyriphos 20 EC @ 25 ml/ kg seed, Pseudomonas maltophilia(PM4) @ 10 ml/kg seed and captan 3 g/kg seed was found to be most effective for control of collar rot and white grub infestation in ground nut.

Selectivity of plant extracts for Trichogramma pretiosum Riley (Hym.: Trichogrammatidae)

We evaluated the selectivity of three plant extracts with potential insecticidal effects for the parasitoid Trichogramma pretiosum Riley, which is commonly used in biological pest control. The plant extracts assayed were an acetone extract of Toona ciliata M. Roem., commercial neem oil, and a nanoencapsulated formulation of neem oil (NC40). The toxicity of the plant extracts to T. pretiosum was evaluated according to the recommendations of the International Organization for Biological Control- IOBC Working Group. We assessed the susceptibility of adults of the maternal and F1 generations and immature stages of T. pretiosum to the extracts. Females exposed to egg cards treated with commercial neem oil parasitized almost 70% fewer eggs than control eggs treated with water; and this extract was therefore classified as slightly harmful. When the eggs were offered to females 24h after treatment with neem oil and aqueous NC40, the parasitism rate also decreased, and the two extracts were classified as slightly harmful. Adult emergence was lower for parasitoids that fed on host eggs offered 24h after the treatment with the T. ciliata extract, which was considered slightly harmful. The emergence of T. pretiosum from eggs, larvae and pupae treated with the different plant extracts, did not decrease compared to development stages treated with the water control. The use of T. pretiosum, combined with the application of an ethanol extract of T. ciliata and a nanoencapsulated formulation of neem, appears to be feasible in view of these low toxicity indices.

Management of the Greater Wax Moth Galleria mellonella with Neem Azal- T/S, in the Laboratory and under Semi-Field Conditions

Abstract Different concentrations of Neem Azal-T/S were used in an artificial diet, to study the mortality of the greater wax moth Galleria mellonella. A Neem formulation and different ages of natural beeswax combs were used for the effective management of the wax moth. While the diet was being prepared, Neem Azal-T/S was directly added ensure that the Neem formulation was distributed evenly in the diet at concentrations of 0.25, 0.5, 1, 2, and 4%. The obtained results showed that the different concentrations of Neem Azal-T/S mixed in the prepared artificial diet had a significant efficacy against the tested 2nd instar larvae. An outstanding elongation of the 2nd larval instar was clear in comparison with the control, even at the least tested concentration (0.25%). Neem Azal-T/S at 4, 2,1, and 0.5% caused 100% mortality for all tested larvae. When using a 4% concentration, all the tested larvae died in the 2nd instar. However, when using a 2% concentration, the larvae died in the fifth instar. When using a decreased concentration of 1.0 and 0.5%, some of the larvae were tolerant and lived till the 6th instar. Feeding the larvae on beeswax combs treated with 2% Neem Azal-T/S, caused 100% mortality when fed on very old wax. When the diet was old wax treated with 2% Neem Azal-T/S, a 91% mortality was recorded. When the diet was new wax treated with 2% Neem Azal-T/S, a 90% mortality was recorded. A 4% Neem formulation caused mortality for all larvae during the first week of treatment on the different tested ages of beeswax combs.